Molecular and Cellular Biochemistry

, Volume 455, Issue 1–2, pp 119–125 | Cite as

Analgesic-antitumor peptide inhibits angiogenesis by suppressing AKT activation in hepatocellular carcinoma

  • Qingxin Cao
  • Wuguang Lu
  • Tingting Zhou
  • Yu Liu
  • Xueting Cai
  • Jin ZhuEmail author
  • Peng CaoEmail author


Hepatocellular carcinoma (HCC) is one of leading causes of cancer-related death, and its increasing incidence worldwide is a cause for concern. The recombinant analgesic-antitumor peptide (rAGAP), a protein consisting of small ubiquitin-related modifier linked with a hexa-histidine tag, exhibited the antitumor activity in HepG2 tumors in our previous study. However, the underlying molecular mechanism of its antitumor activity was still elusive. In this work, we found that treatment with rAGAP reduced phosphorylation of AKT at non-toxic doses in HepG2 cells in vitro. More importantly, treatment of HepG2 cells with rAGAP downregulated protein expression of HIF-1α, suppressed activities of HIF, reduced secretion of VEGF and IL-8, and suppressed HepG2-induced tube formation by HUVEC, which was reversed by co-incubation with SC-79 (an AKT activator). Furthermore, in tumors of athymic mice with HepG2, treatment with rAGAP reduced phosphorylation of AKT, downregulated protein expression of HIF-1α and VEGF, and microvessel density marked by positive CD31 staining. Collectively, rAGAP inhibited angiogenesis by suppressing AKT activation, which partly explained its antitumor activity in HCC.


Recombinant analgesic-antitumor peptide HepG2 Angiogenesis AKT Hepatocellular carcinoma 



Hepatocellular carcinoma


Hypoxia-inducible factor 1 alpha


Human umbilical vein endothelial cells


Interleukin 8


Recombinant antitumor-analgesic peptide


Vascular endothelial growth factor


Author contributions

Conception and design of the experiments: QC, WL, JZ, PC. Collection, analysis, and interpretation of data: QC, WL, TZ, YL, XC, JZ, PC. Drafting the article: QC, WL, JZ, PC.


This study was supported by the project of Quality guarantee system of Chinese Herbal Medicines (201507002), the National Natural Science Foundation of China (81622048, 81473377), China Postdoctoral Science Foundation (2016M601609), and Science Foundation for Distinguished Young Scholars of Jiangsu Province (BK20140049).

Compliance with ethical standards

Conflict of interest

We declare that we have no conflict of interest.

Supplementary material

11010_2018_3475_MOESM1_ESM.docx (494 kb)
Supplementary material 1 (DOCX 493 KB)


  1. 1.
    Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D (2011) Global cancer statistics. CA Cancer J Clin 61:69–90CrossRefGoogle Scholar
  2. 2.
    European Association For The Study of the Liver; European Organisation For Research And Treatment Of Cancer (2012) EASL-EORTC clinical practice guidelines: management of hepatocellular carcinoma. J Hepatol 56:908–943CrossRefGoogle Scholar
  3. 3.
    Omata M, Cheng AL, Kokudo N, Kudo M, Lee JM, Jia J, Tateishi R, Han KH, Chawla YK, Shiina S, Jafri W, Payawal DA, Ohki T, Ogasawara S, Chen PJ, Lesmana CRA, Lesmana LA, Gani RA, Obi S, Dokmeci AK, Sarin SK (2017) Asia–Pacific clinical practice guidelines on the management of hepatocellular carcinoma: a 2017 update. Hepatol Int 11:317–370CrossRefGoogle Scholar
  4. 4.
    Yang JD, Roberts LR (2010) Hepatocellular carcinoma: a global view. Nat Rev Gastroenterol Hepatol 7:448–458CrossRefGoogle Scholar
  5. 5.
    Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A, Schwartz M, Porta C, Zeuzem S, Bolondi L, Greten TF, Galle PR, Seitz JF, Borbath I, Häussinger D, Giannaris T, Shan M, Moscovici M, Voliotis D, Bruix J, SHARP Investigators Study Group (2008) Sorafenib in advanced hepatocellular carcinoma. N Engl J Med 359:378–390CrossRefGoogle Scholar
  6. 6.
    Ding J, Chua PJ, Bay BH, Gopalakrishnakone P (2014) Scorpion venoms as a potential source of novel cancer therapeutic compounds. Exp Biol Med (Maywood) 239:387–393CrossRefGoogle Scholar
  7. 7.
    Chen H, Zhidan W, Xia R, Zhaoxia W, Qing J, Qiang G, Haipeng Y, Hengxiao W (2016) Scorpion venom activates natural killer cells in hepatocellular carcinoma via the NKG2D-MICA pathway. Int Immunopharmacol 35:307–314CrossRefGoogle Scholar
  8. 8.
    Liu X, Liu X, Sunchen S, Liu M, Shen C, Wu J, Zhao W, Yu B, Liu J (2017) A novel tumor-activated ALA fusion protein for specific inhibition on the growth and invasion of breast cancer cells MDA-MB-231. Drug Deliv 24:1811–1817CrossRefGoogle Scholar
  9. 9.
    Al-Asmari AK, Ullah Z, Al Balowi A, Islam M (2017) In vitro determination of the efficacy of scorpion venoms as anti-cancer agents against colorectal cancer cells: a nano-liposomal delivery approach. Int J Nanomedicine 12:559–574CrossRefGoogle Scholar
  10. 10.
    Al Asmari AK, Khan AQ (2016) Investigation of in vivo potential of scorpion venom against skin tumorigenesis in mice via targeting markers associated with cancer development. Drug Des Devel Ther 10:3387–3397CrossRefGoogle Scholar
  11. 11.
    Zhao Y, Cai X, Ye T, Huo J, Liu C, Zhang S, Cao P (2011) Analgesic-antitumor peptide inhibits proliferation and migration of SHG-44 human malignant glioma cells. J Cell Biochem 112:2424–2434CrossRefGoogle Scholar
  12. 12.
    Gu Y, Liu SL, Ju WZ, Li CY, Cao P (2013) Analgesic-antitumor peptide induces apoptosis and inhibits the proliferation of SW480 human colon cancer cells. Oncol Lett 5:483–488CrossRefGoogle Scholar
  13. 13.
    Cao Q, Lu W, Cai X, Hu C, Wang C, Ye J, Yan H, Yang Y, Wang Z, Huo J, Liu Y, Yu Y, Ling C, Cao P (2015) In vitro refolding and functional analysis of polyhistidine-tagged Buthus martensii Karsch antitumor-analgesic peptide produced in Escherichia coli. Biotechnol Lett 37:2461–2466CrossRefGoogle Scholar
  14. 14.
    Ortiz E, Gurrola GB, Schwartz EF, Possani LD (2015) Scorpion venom components as potential candidates for drug development. Toxicon 93:125–135CrossRefGoogle Scholar
  15. 15.
    Lamarca A, Mendiola M, Barriuso J (2016) Hepatocellular carcinoma: Exploring the impact of ethnicity on molecular biology. Crit Rev Oncol Hematol 105:65–72CrossRefGoogle Scholar
  16. 16.
    Yüksel Ş, Boylu Akyerli C, Cengiz Yakıcıer M (2017) Angiogenesis, invasion, and metastasis characteristics of hepatocellular carcinoma. J Gastrointest Cancer. Google Scholar
  17. 17.
    Ghouri YA, Mian I, Rowe JH (2017) Review of hepatocellular carcinoma: epidemiology, etiology, and carcinogenesis. J Carcinog 16:1CrossRefGoogle Scholar
  18. 18.
    Luo D, Wang Z, Wu J, Jiang C, Wu J (2014) The role of hypoxia inducible factor-1 in hepatocellular carcinoma. Biomed Res Int 2014:409272Google Scholar
  19. 19.
    Ferrara N, Gerber HP, LeCouter J (2003) The biology of VEGF and its receptors. Nature Med 9:669–676CrossRefGoogle Scholar
  20. 20.
    Courtney KD, Corcoran RB, Engelman JA (2010) The PI3K pathway as drug target in human cancer. J Clin Oncol 28:1075–1083CrossRefGoogle Scholar
  21. 21.
    Li W, Tan D, Zhang Z, Liang JJ, Brown RE (2008) Activation of Akt-mTOR-p70S6K pathway in angiogenesis in hepatocellular carcinoma. Oncol Rep 20:713–719Google Scholar
  22. 22.
    Gao F, Li H, Chen YD, Yu XN, Wang R, Chen XL (2009) Upregulation of PTEN involved in scorpion venom-induced apoptosis in a lymphoma cell line. Leuk Lymphoma 50:633–641CrossRefGoogle Scholar
  23. 23.
    Du J, Wang R, Yin L, Fu Y, Cai Y, Zhang Z, Liang A (2018) B < i> m</i> K CT enhances the sensitivity of temozolomide-induced apoptosis of malignant glioma U251 cells < i> in vitro</i> through blocking the AKT signaling pathway. Oncol Lett 15:1537–1544.Google Scholar
  24. 24.
    Chai ZT, Kong J, Zhu XD, Zhang YY, Lu L, Zhou JM, Wang LR, Zhang KZ, Zhang QB, Ao JY, Wang M, Wu WZ, Wang L, Tang ZY, Sun HC (2013) MicroRNA-26a inhibits angiogenesis by down-regulating VEGFA through the PIK3C2α/Akt/HIF-1α pathway in hepatocellular carcinoma. PLoS ONE 8:e77957CrossRefGoogle Scholar
  25. 25.
    Lau CK, Yang ZF, Ho DW, Ng MN, Yeoh GC, Poon RT, Fan ST (2009) An Akt/hypoxia-inducible factor-1alpha/platelet-derived growth factor-BB autocrine loop mediates hypoxia-induced chemoresistance in liver cancer cells and tumorigenic hepatic progenitor cells. Clin Cancer Res 15:3462–3471CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Huadong Medical Institute of BiotechniquesNanjingPeople’s Republic of China
  2. 2.Affiliated Hospital of Integrated Traditional Chinese and Western MedicineNanjing University of Chinese MedicineNanjingPeople’s Republic of China
  3. 3.Laboratory of Cellular and Molecular BiologyJiangsu Province Academy of Traditional Chinese MedicineNanjingPeople’s Republic of China
  4. 4.Drum Tower HospitalNanjingPeople’s Republic of China

Personalised recommendations